In vitro expansion and genetic modification of gastrointestinal stem cells in spheroid culture

Nature Protocols

Volumn 8, Issue 12, 2013, Pages 2471-2482

  Miyoshi, Hiroyuki ^a   Stappenbeck, Thaddeus S ^a  

^a WASHINGTON UNIVERSITY SCHOOL OF MEDICINE   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

MATRIGEL;

ANIMAL CELL; ANIMAL EXPERIMENT; ANIMAL TISSUE; ARTICLE; CELL ASSAY; CELL ISOLATION; CELL SELECTION; CELL VIABILITY; CRYOPRESERVATION; DNA MODIFICATION; EPITHELIUM CELL; FEMALE; GASTROINTESTINAL STEM CELL; GASTROINTESTINAL TRACT; IN VITRO STUDY; INFECTION RATE; LENTIVIRINAE; MALE; MICROBIAL CONTAMINATION; MOUSE; NONHUMAN; PIPETTE; PRIORITY JOURNAL; SPHEROID CELL; STEM CELL; STEM CELL EXPANSION; STROMA CELL;

MUS;

EID: 84888331271     PISSN: 17542189     EISSN: 17502799     Source Type: Journal    
DOI: 10.1038/nprot.2013.153     Document Type: Article

References (14)

1. Sato, T. et al. Single Lgr5 stem cells build crypt-villus structures in vitro without a mesenchymal niche. Nature 459, 262-266 (2009).
2. +ve stem cells drive self-renewal in the stomach and build long-lived gastric units in vitro. Cell Stem Cell 6, 25-36 (2010).
3. Miyoshi, H., Ajima, R., Luo, C.T., Yamaguchi, T.P. & Stappenbeck, T.S. Wnt5a potentiates TGF-β signaling to promote colonic crypt regeneration after tissue injury. Science 338, 108-113 (2012).
4. Nusse, R. et al. Wnt signaling and stem cell control. Cold Spring Harb. Symp. Quant. Biol. 73, 59-66 (2008).
5. Haegebarth, A. & Clevers, H. Wnt signaling, Lgr5, and stem cells in the intestine and skin. Am. J. Pathol. 174, 715-721 (2009).
6. Kim, K.A. et al. Mitogenic influence of human R-spondin1 on the intestinal epithelium. Science 309, 1256-1259 (2005).
7. Willert, K. et al. Wnt proteins are lipid-modified and can act as stem cell growth factors. Nature 423, 448-452 (2003).
8. Barker, N. et al. Identification of stem cells in small intestine and colon by marker gene Lgr5. Nature 449, 1003-1007 (2007).
9. Watanabe, K. et al. A ROCK inhibitor permits survival of dissociated human embryonic stem cells. Nat. Biotechnol. 25, 681-686 (2007).
10. Koo, B.K. et al. Controlled gene expression in primary Lgr5 organoid cultures. Nat. Methods 9, 81-83 (2011).
11. Nagy, A. Manipulating the Mouse Embryo: A Laboratory Manual. (Cold Spring Harbor Laboratory Press, 2003).
12. Sato, T. et al. Long-term expansion of epithelial organoids from human colon, adenoma, adenocarcinoma, and Barrett's epithelium. Gastroenterology 141, 1762-1772 (2011).
13. Sato, T. Novel stem cell culture system. Inflamm. Regen. 32, 43-47 (2012).
14. Snippert, H.J. et al. Intestinal crypt homeostasis results from neutral competition between symmetrically dividing Lgr5 stem cells. Cell 143, 134-144 (2010).